miR-let-7a调节PI3K/Akt信号通路抑制人系膜细胞增殖和炎症反应  

The role and mechanism of miR-let-7a in regulating PI3K/Akt signaling pathway to inhibit human mesangial cell proliferation and inflammation

作　　者：罗宇茜 高悦 叶晓坤 王安琪 韩迎雯 邵晓轶 LUO Yu-xi;GAO Yue;YE Xiao-kun;WANG An-qi;HAN Ying-wen;SHAO Xiao-yi(Department of Immunology,Nantong University Medical School,Nantong 226001,China)

机构地区：[1]南通大学医学院免疫学系,南通226001

出　　处：《现代免疫学》2023年第6期449-455,464,共8页Current Immunology

基　　金：国家自然科学基金面上项目(81771767);国家自然科学基金青年科学基金项目(81801580)。

摘　　要：为探究miR-let-7a对LPS诱导的人系膜细胞(human mesangial cell,HMC)增殖和炎症反应的影响及其通过调节PI3K/Akt信号通路发挥作用的机制,体外条件下用0.1μg/mL LPS处理HMC,CCK-8法检测各时间点细胞活性;Western blotting检测HMC增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、凋亡相关蛋白BAX及信号通路蛋白PI3K、p-Akt的表达情况;qPCR检测细胞中miR-let-7a及炎性因子TNF-α、IL-1β、IL-6和Ⅳ型胶原蛋白mRNA的表达。转染miR-let-7a mimic或inhibitor改变HMC中miR-let-7a的表达,或使用抑制剂LY294002阻断HMC中PI3K/Akt信号通路,观察不同处理对HMC增殖、炎症反应及PI3K/Akt信号通路激活的影响。结果显示,LPS处理可诱导HMC增殖和炎症反应,抑制miR-let-7a的表达(均P<0.05)并激活PI3K/Akt信号通路(均P<0.01);与LPS组相比,过表达miR-let-7a可下调LPS诱导的HMC增殖和炎症反应,抑制PI3K/Akt信号通路的激活(均P<0.05);反之,下调miR-let-7a水平可促进由LPS刺激引起的细胞增殖、炎症反应及PI3K/Akt信号通路激活(均P<0.05);阻断PI3K/Akt信号通路也可抑制HMC的活性和炎性因子产生(均P<0.05)。综上,在经LPS处理的HMC中,miR-let-7a可通过调节PI3K/Akt信号通路抑制HMC的增殖与炎症反应。The purpose of this study was to investigate the effect of miR-let-7a on the proliferation and inflammatory response of human mesangial cell(HMC)induced by LPS and the mechanism by which miR-let-7a acts in regulating PI3K/Akt signaling pathway.To this end,HMCs were treated with 0.1μg/mL LPS in vitro.The cell viability was measured by CCK-8 assay;the expressions of proliferating cell nuclear antigen(PCNA),apoptosis-related protein BAX,and signaling pathway proteins PI3K and p-Akt were detected by Western blotting;the mRNA levels of miR-let-7a and inflammatory factors TNF-α,IL-1β,IL-6 and collagenⅣin HMC were measured by qPCR.The expression of miR-let-7a in HMC was motified by transfecting either miR-let-7a mimic or inhibitor and the PI3K/Akt signaling pathway was blocked by using the inhibitor LY294002.The effects of the above treatments on HMC proliferation,inflammatory response,and the activation of PI3K/Akt signaling pathway were examined.The results showed that LPS stimulation induced the proliferation and inflammatory response of HMC,inhibited the expression of miR-let-7a(all P<0.05),and activated the PI3K/Akt signaling pathway(all P<0.01).Compared to LPS-treated group,over-expression of miR-let-7a down-regulated LPS-induced HMC proliferation and inflammatory response and inhibited the activation of the PI3K/Akt signaling pathway(all P<0.05).On the other hand,down-regulation of miR-let-7a promoted cell proliferation,inflammatory response,and PI3K/Akt signaling pathway activation induced by LPS stimulation(all P<0.05).Blocking the PI3K/Akt signaling pathway also inhibited HMC viability and the production of inflammatory factors(all P<0.05).In conclusion,in LPS-stimulated HMC,miR-let-7a inhibits the proliferation and inflammatory response of HMC by regulating the PI3K/Akt signaling pathway.

关 键 词：脂多糖 人系膜细胞 微小核糖核酸let-7a 磷脂酰肌醇3激酶/蛋白激酶B信号通路 细胞增殖 

分 类 号：R392.12[医药卫生—免疫学]